Berkeley LBL/JoyceNotebook

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'''Contruction of pET3a Derivatives Containing ''bchD'' and ''bchI'''''
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[[User:Joyxi|'''Joyce's user page]]
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1. Amplify Rhodobacter's gene and introduce restriction sites by [[Berkeley_LBL/PCRphusion|PCR (Using Phusion Polymerase)]], which allow cloning PCR fragments into pET3a
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[[Berkeley_LBL/JoyceRhodobacter|'''Contruction of pET3a Derivatives Containing ''bchD'' and ''bchI''''']]
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{| border =
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!
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!''bchD''  
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!''bchI''
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[[Berkeley_LBL/JoyceCyano|'''Contruction of pET3a Derivatives Containing ''chlD'' and ''chlI''''']]
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|length of gene fragment
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|1695 b.p.
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|1005 b.p.
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|Restriction sites introduced when amplified by PCR
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|NdeI, BamHI
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|NdeI, BglI
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2. Do [[Berkeley_LBL/Digestion2|Analytic Digestion]] to confirm that the length of the PCR fragments are the same as the corresponding target gene fragments.
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3. Remove any leftover PCR enzyme in the samples by [[Berkeley_LBL/PCRcleanup|PCR Clean Up/Purification]].
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4. [[Berkeley_LBL/Digestion|Digestion for PCR Products]] using specific restriction enzymes
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{| border =
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!''bchD''  
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|NdeI, BamHI
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!''bchI''
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|NdeI, BglI
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|}
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Latest revision as of 02:35, 27 October 2007

Joyce's user page

Contruction of pET3a Derivatives Containing bchD and bchI

Contruction of pET3a Derivatives Containing chlD and chlI

Retrieved from "http://2007.igem.org/wiki/index.php/Berkeley_LBL/JoyceNotebook"